Inflammasome Activation Triggers Systemic Coagulation in Sepsis

脓毒症中炎症小体激活引发全身凝血

• 批准号: 10645452
• 负责人: ZHENYU Li
• 金额: $ 42.95万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10645452
• 关键词: Apoptosis; Bacteria; Bacterial Infections; Blood Coagulation Disorders; Blood Coagulation Factor; Bone Marrow; Burkholderia; CASP1 gene; Cells; Cessation of life; Clinical; Coagulation Process; Complication; Consumption; Data; Development; Disease; Disseminated Intravascular Coagulation; Escherichia coli; Family; Fibrinogen; Flagellin; Functional disorder; Goals; Gram-Negative Bacteria; In Vitro; Infection; Inflammasome; Infusion procedures; Injections; Intervention; Knockout Mice; Lead; Link; Modeling; Molecular; Morbidity - disease rate; Mus; Myelogenous; Needles; Organ; Outcome; Pathway interactions; Patients; Peripheral; Phagocytosis; Physiological; Plasma; Play; Proteins; Prothrombin time assay; Pseudomonas aeruginosa; Reagent; Research Project Grants; Rod; Role; Salmonella; Sepsis; Septic Shock; Testing; Thrombocytopenia; Thromboplastin; Thrombosis; Thrombus; Tissues; Type III Secretion System Pathway; Work; antithrombin III-protease complex; base; cecal ligation puncture; driving force; in vivo; insight; intravenous injection; macrophage; monocyte; mortality; mouse model; multidisciplinary; new therapeutic target; protein E; release factor; septic patients; targeted treatment

Abstract Septic shock is invariably associated with systemic coagulation leading to thrombus formation. Sepsis-related organ dysfunction has been attributed to microvascular thrombosis. Mortality rate doubles in septic patients with disseminated intravascular coagulation (DIC). DIC is even considered as a sign that “death is coming”. Previous studies have demonstrated the important roles of tissue factor (TF) in sepsis-associated DIC. However, the mechanism leading to TF release, which triggers systemic coagulation in sepsis, is unknown. Recent in vitro studies revealed that bacterial components (flagellin, the rod protein of the type III secretion system (T3SS), or LPS) induce programmed cell death (called pyroptosis) of macrophages through activation of inflammasome pathways. We show that intravenous injection of a T3SS rod protein E. coli, EprJ, induced depletion of peripheral monocytes and macrophages in tissues. Importantly, injection of EprJ or LPS, which elicit canonical and noncanonical inflammasome activation, respectively, induced systemic coagulation activation, as evident by prolonged prothrombin time (PT) due to increased consumption of coagulation factors, thrombocytopenia, increased plasma levels of thrombin-antithrombin complex (TAT), and reduced plasma fibrinogen levels. Thus, our findings made connections between the in vitro and in vivo observations and suggested monocyte/macrophage pyroptosis as a trigger of DIC in sepsis. The goal of this application is to delineate the underlying mechanisms by which inflammasome activation and pyroptosis trigger DIC in sepsis. Specific Aim 1 will establish inflammasome activation and pyroptosis as a common mechanism for DIC induced by bacterial infection. The working hypothesis is that bacteria and bacterial components from different strains elicit DIC through Inflammasome activation and pyroptosis. We will use a combination of various deficient mice to elucidate the role of inflammasome activation and pyroptosis in DIC elicited by Gram-negative bacteria. Specific Aim 2 is to identify the molecular mechanism of TF release from macrophages following inflammasome activation. We will also use the myeloid-specific TF knockout mice and a low TF mouse model to elucidate whether DIC elicited by the bacterial components depends on release of TF from macrophages. Specific Aim 3 will demonstrate the role of inflammasome activation in sepsis-associated coagulopathy. We will use the cecal ligation and puncture (CLP) sepsis model and bacterial infusion sepsis model to investigate the role of inflammasome activation and pyroptosis in coagulation. Completion of the proposed studies will demonstrate the molecular mechanism of systemic coagulation is sepsis. Such findings would have profound ramifications for the identification of new drug targets for DIC, the deadly complication of sepsis.

摘要 感染性休克总是与导致血栓形成的全身凝血有关。感染相关 器官功能障碍归因于微血管血栓形成。败血症患者死亡率翻倍 弥散性血管内凝血（DIC）DIC甚至被认为是“死亡即将来临”的征兆。 以往的研究表明，组织因子（TF）在脓毒症相关DIC中的重要作用。 然而，导致TF释放的机制尚不清楚，TF释放触发脓毒症中的全身凝血。 最近的体外研究表明，细菌组分（鞭毛蛋白，III型分泌的杆状蛋白） 系统（T3 SS）或LPS）通过激活诱导巨噬细胞程序性细胞死亡（称为焦亡 炎性体通路。我们发现，静脉注射T3 SS杆蛋白E。大肠杆菌，EprJ，诱导 组织中的外周单核细胞和巨噬细胞耗竭。重要的是，注射EprJ或LPS， 分别引起典型和非典型炎性小体激活，诱导全身凝血 活化，如凝血因子消耗增加导致凝血酶原时间（PT）延长所证明， 血小板减少症，凝血酶-抗凝血酶复合物（达特）的血浆水平升高， 纤维蛋白原水平。因此，我们的研究结果在体外和体内观察之间建立了联系， 提示单核细胞/巨噬细胞凋亡是败血症DIC的触发因素。此应用程序的目标是 阐明炎症小体激活和焦亡触发脓毒症DIC的潜在机制。 特异性目的1将确定炎性小体激活和焦亡是DIC的常见机制 由细菌感染引起的。工作假设是，细菌和细菌成分来自不同的 菌株通过炎性小体激活和焦亡引起DIC。我们将结合使用各种 缺陷小鼠，以阐明炎症小体激活和焦亡在革兰氏阴性杆菌引起的DIC中的作用。 细菌具体目标2是确定巨噬细胞释放TF的分子机制， 炎性小体激活。我们还将使用骨髓特异性TF敲除小鼠和低TF小鼠模型 阐明细菌成分引起的DIC是否依赖于巨噬细胞释放TF。 特异性目的3将证明炎性小体活化在脓毒症相关凝血病中的作用。我们将 采用盲肠结扎穿孔（CLP）脓毒症模型和细菌输注脓毒症模型， 炎性小体活化和焦亡在凝血中的作用。完成拟议的研究将 证明全身凝血的分子机制是脓毒症。这样的发现将对 为确定DIC（败血症的致命并发症）的新药靶点提供了新的思路。

项目成果

Inhibiting NINJ1-dependent plasma membrane rupture protects against inflammasome-induced blood coagulation and inflammation.

抑制 NINJ1 依赖性质膜破裂可防止炎症体诱导的凝血和炎症。

• DOI: 10.1101/2023.08.30.555561
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Cui,Jian;Li,Hua;Zhang,Guoying;Zhang,Yan;Yang,Ling;Sim,MarthaMS;Wood,JeremyP;Wei,Yinan;Li,Zhenyu;Wu,Congqing
• 通讯作者: Wu,Congqing

Inflammasome activation and pyroptosis mediate coagulopathy and inflammation in Salmonella systemic infection.

• DOI: 10.1016/j.micres.2023.127460
• 发表时间: 2023-10
• 期刊: MICROBIOLOGICAL RESEARCH
• 影响因子: 6.7
• 作者: Pandeya, Ankit;Zhang, Yan;Cui, Jian;Yang, Ling;Li, Jeffery;Zhang, Guoying;Wu, Congqing;Li, Zhenyu;Wei, Yinan
• 通讯作者: Wei, Yinan

Calcium Ion Chelation Preserves Platelet Function During Cold Storage.

• DOI: 10.1161/atvbaha.120.314879
• 发表时间: 2021-01
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Xiang B;Zhang G;Zhang Y;Wu C;Joshi S;Morris AJ;Ware J;Smyth SS;Whiteheart SW;Li Z
• 通讯作者: Li Z